Luteal Research Articles

Selecting optimal progestational agents either alone or in combination in common pediatric endocrine settings: challenges of unmet needs.

Progesterone is a natural hormone, mainly produced by the corpus luteum, with the foremost endocrine function on the secretory glands of the endometrium. Since being isolated, both natural and synthetic forms have been produced and are utilized for several purposes, including regulating the menstrual cycle and preventing endometrial hyperplasia. Specifically, the use of progestational agents is essential in the treatment of many common endocrine conditions, including Polycystic Ovary Syndrome, Congenital Adrenal Hyperplasia, Turner Syndrome, and Functional Hypothalamic Amenorrhea. Although these agents are essential for disease management, literature that focuses on the benefits of specific progestins as well as the effects on glucocorticoid receptors (GR), mineralocorticoid receptors (MR), and androgenic receptors (AR) is limited. In this review, we provide a disease specific summary of the available literature and highlight where more information is needed.

The Potential Health Risks of Catha edulis Forsk (Khat) Consumption on Some Vital Organs

Abstract Introduction/Objective Introduction: Catha edulis Forsk (Khat) leaves are widely chewed by adults in the Arabian Peninsula and the Horn of Africa for their mild stimulating properties. Objectives To investigate the impact of an aqueous extract of Catha edulis Forsk leaves on the proper functioning of vital organs. Methods/Case Report Methods: Materials and methods: Twenty-five female albino rats were divided into two groups: a control group and a treated group which received a daily gavage of 1 ml of aqueous Khat extract with a concentration of 700 mg/kg body weight for four weeks. The effects of this treatment on selected organs, including the brain, hippocampus, heart, and ovaries, were assessed using various measurements and analyses. Results (if a Case Study enter NA) Results: Results: The antioxidant evaluation showed a significant increase in MDA levels in the khat group and a significant decrease in GSH, SOD, and CAT levels compared to the control group. There was a significant decrease in IL-4 and a significant increase in TNF-α and TGF-β1 compared to the control group. In contrast, the cardiac enzymes; Troponin, CK, CPK, and GOT showed a significant increase compared to the control group. The results of the hormonal assay indicated a significant increase in LH, FSH, and testosterone levels, along with a significant decrease in estrogen in the khat group compared to the control group. Furthermore, the histological investigations revealed significant changes in tissue architecture in the studied organs. In the heart, the treated group exhibited an intact, thick pericardium, along with markedly apoptotic cardiac muscle fibers and others with pyknotic nuclei and mildly congested blood vessels and capillaries. The examination of brain tissues in the treated group showed thick meninges with mild inflammatory infiltrate, marked astrogliosis with degenerated neurons in the cerebral cortex, inflammatory infiltrate with abscess formation and mildly congested blood vessels and eosinophilic plaque-like area in the deep cortex, and scattered degenerated neurons and some neurons with nuclear vacuoles and average glial cells in the white matter. In the ovaries, sections showed primary, secondary, and antral follicles, scattered cystically dilated follicles with atrophied lining, and multiple corpora lutea in the edematous stroma with scattered apoptotic stromal cells. Conclusion Based on these findings, long-term use of khat as a chewing material may harm vital organs.

Genetic architecture of meat traits in Large White sows

Currently, genome-wide association analysis is a modern and reliable method for analyzing genomic information about animals, as well as determining the “genotype — phenotype” relationship. This study aims to use the GWAS method to identify significant SNPs located within or linked to genes for meat traits in Large White pigs — backfat thickness over the 6–7th and 10–12th vertebrae, and loin muscle depth. The conducted GWA analysis revealed 60 genes, of which 17 are associated with biological functionality, annotated using the DAVID program. Three genes were found to have codification in the Pig QTL database. The genes were divided into 10 groups based on gene ontology (GO). Of all the genes, the AUTS2 gene, located on chromosome 3 and predicting the number of corpora lutea in sows, is of greatest interest. The results of this scientific work will contribute to the development of a genetic evaluation system and improvement of meat qualities in pigs.

Small extracellular vesicles derived from the crosstalk between early embryos and the endometrium potentially mediate corpus luteum function.

The first interactions among the embryo, endometrium, and corpus luteum (CL) are essential for pregnancy success. Small extracellular vesicles (sEVs) are part of these interactions. We previously demonstrated that sEVs from in vivo- or in vitro-produced bovine embryos contain different miRNA cargos. Herein we show: 1) the presence and origin (in vivo or in vitro) of the blastocyst differentially reprograms endometrial transcriptional profiles; 2) the endometrial explant (EE) cultured with in vivo or in vitro embryos release sEVs with different miRNA contents, and; 3) the luteal explant (CLE) exposed to these sEVs have distinct mRNA and miRNA profiles. To elucidate this, the EE were cultured in the presence or absence of a single Day-7 in vivo (EE-AI) or in vitro (EE-IVF) embryo. After of culture we found, in the EE, 45 and 211 differentially expressed genes (DEGs) associated with embryo presence and origin, respectively. SEVs were recovered from the conditioned media (CM) in which EE and embryos were co-cultured. Four miRNAs were differentially expressed between sEVs from CM-EE-AI and CM-EE-IVF. Luteal explants exposed in culture to these sEVs showed 1360 transcripts, and fifteen miRNAs differentially expressed. The DEGs associated with embryo presence and origin, modulating cells' proliferation, and survival. These results demonstrate that in vivo- or in vitro-produced bovine embryos induce molecular alterations in the endometrium; and that the embryo and endometrium release sEVs capable of modifying the mRNA and miRNA profile in the CL. Therefore, the sEVs-mediated embryo-endometrium-CL interactions possibly regulate the CL viability to ensure pregnancy success.

Fertility and teratogenic study of the aqueous extract of Hibiscus sabdariffa L. in female Sprague Dawley rats

Hibiscus sabdariffa (roselle) has long been gaining attention in phytotherapy research due to its medicinal properties. Despite its benefits, there is very scarce information available on the reproductive toxicity of this plant. The aqueous extract of H. sabdariffa (AEHS) was tested in the present study to investigate its potential effects on fertility and teratogenicity in 40 female Sprague Dawley rats. AESH treatment was administered orally by gavage at four different dosages: 250, 500 and 1000 mg/kg/day or distilled water (control). Treatment began from pre-mating and continued through mating up to the 19th day of pregnancy periods. Throughout this study, the reproductive parameters were evaluated until the day of sacrifice (day 20th of pregnancy). Results obtained revealed no significant differences in general physical health, behaviours and maternal body weights throughout the treatment period. Furthermore, the mean length of the oestrous cycle was not statistically affected, even though a few rats displayed irregular cycles. In addition, there was no statistical significance in the mating and pregnancy indices, number of corpora lutea and implantation sites, percentages of pre-implantation loss and postimplantation death and reproductive organ weights. Foetal parameters such as the number of live foetuses, sex ratio and body weight were also not statistically affected by AESH. Ultimately, there were no signs of teratogenicity observed since none of the foetuses exhibited congenital malformations. In conclusion, these findings imply that the oral administration of AEHS up to 1000 mg/kg/day did not pose any significant toxicity on the fertility and teratogenicity but slightly affected the oestrous cycle in rats.

Antioxidant activity of self-nanoemulsifying drug delivery system (SNEDDS) of Curcuma longa in polycystic ovary syndrome rat model

Purpose: To investigate the effect of SNEDDS containing Curcuma longa extract on hormonal and histological improvement of polycystic ovary syndrome (PCOS) rat model. Method: A total of 36 female Wistar rats, aged between 8 and 10 weeks, were randomly divided into 6 groups, including control (normal rats), PCOS (untreated PCOS rats), PCOS + M (treated with 20 mg/kg bw/day metformin), PCOS + SC25, PCOS + SC50, and PCOS + SC100 (treated with Curcuma longa extract at 25, 50, and 100 mg/kg/day with SNEDDS, respectively). This study used a posttest-only control group design. The rats were euthanized on day 15, and blood samples were taken to examine malondialdehyde (MDA) and anti-mullerian hormone (AMH) levels. Ovarian tissue was prepared on a histological slide, and various follicles were observed. Data was analyzed using analysis of variance (ANOVA) and Kruskal-Wallis tests. Results: Curcuma longa extract, at 50 and 100 mg/kg, incorporated in SNEDDS resulted in a significant reduction in the corpus luteum number and width of the granulosa layer (p < 0.05). Also, Curcuma longa significantly reduced malondialdehyde (MDA), anti-mullerian hormone (AMH), preantral follicles, and follicular cysts (p < 0.05). Conclusion: Curcuma longa extract in SNEDDS reduces MDA and AMH levels and improves the histology of the ovaries in PCOS model rats. There is a need to conduct further studies of C. longa extract or isolates in SNEDDS using human cell lines of PCOS model.

8354 OVARIAN HYPERTHECOSIS: AN UNCOMMON CAUSE OF HYPERANDROGENISM IN A POSTMENOPAUSAL WOMAN

Abstract Disclosure: M.D. Severino: None. F.P. Silva: None. M.V. Lopes: None. C. Nunes: None. M.I. Alexandre: None. M.J. Bugalho: None. Introduction: Severe hirsutism in postmenopausal women demands a careful clinical evaluation to exclude a malignant cause, such as adrenal or ovarian androgen-secreting tumors. Ovarian hyperthecosis is an uncommon and benign cause of hyperandrogenism and, consequently, hirsutism. It is caused by persistent testosterone synthesis due to proliferation of ovarian stromal cells and nests of luteinized theca cells. It occurs more often in postmenopausal women, as aromatization of testosterone to estradiol decreases due to the loss of granulosa cells. It is also frequently associated with obesity and insulin resistance. Clinical case: We present the case of a 61-year-old woman, with a history of hypertension and dyslipidemia, referred to Endocrinology Clinic due to a year long history of hirsutism, most severe on her face, chest, and abdomen. She also mentioned slight clitoromegaly. She had no other virilization symptoms such as acne, male-pattern alopecia, voice deepening or mammary atrophy. Her menarche occurred at the age of 14 and she had regular menses until menopause, at the age of 51. She had a BMI of 32 kg/m2 and an uncharacteristic phenotype. At observation, she had a mild hirsutism (Ferriman-Gallwey score of 9). This contrasted with the biochemical findings of severe androgenism - total testosterone of 366 ng/dL (reference values [RV] for women between 5-48) with free testosterone 7.8 pg/mL and SHBG 74 nmol/L (RV 32-128), resulting in a Free Androgen Index of 17,16 (RV <6.6). DHEAS, androstenedione, 17-hydroxyprogesterone and prolactin were normal, and gonadotropins were in the menopausal range. Blood tests also showed hyperinsulinemia and pre-diabetes with HbA1c 6,2%. She denied exposure to steroids in the form of new medication, supplements, or creams/gels (including second-hand exposure through her partner) and she had no exposure to other drugs with androgenic effect. Abdominal and pelvic CT scan, as well as transvaginal ultrasound, showed no adrenal or ovarian lesions. An ovarian cause was deemed the most likely and laparoscopic diagnostic bilateral salpingo-oophorectomy was performed. Gross pathology revealed ovaries of regular size, while histology showed bilateral ovarian stromal hyperthecosis. Testosterone levels returned to normal one month after surgery (14,2 ng/dL). Conclusion: Benign, ovarian causes of hyperandrogenism are uncommon in postmenopausal women, requiring a high index of suspicion. The diagnosis is difficult, and laparoscopic bilateral salpingo-oophorectomy is essential as both a diagnostic and therapeutic tool. Presentation: 6/2/2024

8354 Ovarian Hyperthecosis: an Uncommon Cause of Hyperandrogenism in a Postmenopausal Woman

Abstract Disclosure: M.D. Severino: None. F.P. Silva: None. M.V. Lopes: None. C. Nunes: None. M.I. Alexandre: None. M.J. Bugalho: None. Introduction: Severe hirsutism in postmenopausal women demands a careful clinical evaluation to exclude a malignant cause, such as adrenal or ovarian androgen-secreting tumors. Ovarian hyperthecosis is an uncommon and benign cause of hyperandrogenism and, consequently, hirsutism. It is caused by persistent testosterone synthesis due to proliferation of ovarian stromal cells and nests of luteinized theca cells. It occurs more often in postmenopausal women, as aromatization of testosterone to estradiol decreases due to the loss of granulosa cells. It is also frequently associated with obesity and insulin resistance. Clinical case: We present the case of a 61-year-old woman, with a history of hypertension and dyslipidemia, referred to Endocrinology Clinic due to a year long history of hirsutism, most severe on her face, chest, and abdomen. She also mentioned slight clitoromegaly. She had no other virilization symptoms such as acne, male-pattern alopecia, voice deepening or mammary atrophy. Her menarche occurred at the age of 14 and she had regular menses until menopause, at the age of 51. She had a BMI of 32 kg/m2 and an uncharacteristic phenotype. At observation, she had a mild hirsutism (Ferriman-Gallwey score of 9). This contrasted with the biochemical findings of severe androgenism - total testosterone of 366 ng/dL (reference values [RV] for women between 5-48) with free testosterone 7.8 pg/mL and SHBG 74 nmol/L (RV 32-128), resulting in a Free Androgen Index of 17,16 (RV <6.6). DHEAS, androstenedione, 17-hydroxyprogesterone and prolactin were normal, and gonadotropins were in the menopausal range. Blood tests also showed hyperinsulinemia and pre-diabetes with HbA1c 6,2%. She denied exposure to steroids in the form of new medication, supplements, or creams/gels (including second-hand exposure through her partner) and she had no exposure to other drugs with androgenic effect. Abdominal and pelvic CT scan, as well as transvaginal ultrasound, showed no adrenal or ovarian lesions. An ovarian cause was deemed the most likely and laparoscopic diagnostic bilateral salpingo-oophorectomy was performed. Gross pathology revealed ovaries of regular size, while histology showed bilateral ovarian stromal hyperthecosis. Testosterone levels returned to normal one month after surgery (14,2 ng/dL). Conclusion: Benign, ovarian causes of hyperandrogenism are uncommon in postmenopausal women, requiring a high index of suspicion. The diagnosis is difficult, and laparoscopic bilateral salpingo-oophorectomy is essential as both a diagnostic and therapeutic tool. Presentation: 6/2/2024

7617 Two Cases of Severe Beta-hCG-Mediated Hyperthyroidism with Gestational Trophoblastic Disease

Abstract Disclosure: A. Morvant: None. M. Mashayekhi: None. Introduction: Hyperthyroidism is most commonly caused by Graves’ disease and toxic nodules and affects 2.5% of the world’s population. Hyperthyroidism due to elevations in beta-hCG (b-hCG), which has thyroid-stimulating activity due to homology with TSH, is a rare process that requires high clinical suspicion. We present two cases of hyperthyroidism caused by severely elevated b-hCG due to gestational trophoblastic disease (GTD), with rapid resolution of hyperthyroidism after treatment of the underlying conditions. Clinical Cases Case 1: A 35-year-old at 28-weeks gestation presented with dyspnea, palpitations, and 6-pound weight loss over the prior month. Labs revealed b-hCG 5.2 million (0-5 mIU/mL), TSH <0.015 (0.35-3.6 mcU/mL), and free T4 2.42 (0.7-1.37 ng/dL). Imaging revealed extensive pulmonary lesions concerning for metastatic disease and a 2.4 cm ovarian mass. Due to worsening respiratory distress and the need to start chemotherapy urgently, she underwent Cesarean section to deliver the viable infant and take out the ovarian mass. Pathology confirmed choriocarcinoma. Methimazole was started post-operatively, and she started chemotherapy. Labs after cycle one of chemotherapy showed b-hCG 1.3 million, TSH <0.015, and free T4 1.46. The methimazole dose was decreased and ultimately stopped as free T4 levels decreased with subsequent chemotherapy cycles. After three cycles, labs showed b-hCG 4,652, TSH 1.55, and free T4 0.68. She was discharged without thyroid-targeting therapy. Case 2: A 33-year-old woman at 12-weeks gestation presented with nausea, vomiting, and 10-pound weight loss over 1 month. Labs showed TSH <0.015, free T4 2.39, and total T3 351 (58-160 ng/dL), which were thought to be due to hyperemesis gravidarum. Two months later she presented with vaginal bleeding and imaging showed an enlarged placenta with a moth-eaten appearance and theca lutein cysts concerning for partial molar pregnancy. Beta-hCG level was 3.3 million, TSH <0.015, free T4 1.76, and total T3 289. She underwent dilation and evacuation and pathology confirmed partial hydatidiform mole. Beta-hCG level decreased to 1.8 million and free T4 normalized after surgery, with b-hCG 3,300, normal TSH, and normal total T3 one month later. Conclusion: These cases highlight the importance of considering possible GTD when evaluating a patient with hyperthyroidism, particularly in women of childbearing age and cases of early pregnancy when the degree of hyperthyroidism seems out of proportion to that expected. Clinicians should consider checking a b-hCG level in consultation with an obstetrician when indicated. Treatment is aimed at the GTD and can yield rapid resolution of hyperthyroidism.

7869 Conditional deletion of androgen receptors from kisspeptin neurons only partially rescues the reproductive phenotype of a prenatal androgen-induced mouse model of polycystic ovary syndrome

Abstract Disclosure: A.G. Novak: None. A.V. Petriv: None. D.B. Lohr: None. K.M. Dillon: None. N.T. Neifert: None. A.M. Moore: None. Polycystic ovary syndrome (PCOS) is a prevalent cause of infertility in people with ovaries of reproductive age. Although the etiology of PCOS remains unclear, research in the clinic and with animal models suggests that hyperandrogenism (the presence of elevated circulating androgens) plays a significant role in the development of reproductive dysfunction, including irregular menstrual/estrous cycles and altered ovarian morphology. Although the site at which androgens disrupt the reproductive axis is unclear, neurons expressing kisspeptin are crucial regulators of the hypothalamic-pituitary-gonadal axis and express androgen receptors. Therefore, we hypothesize that androgen signaling at kisspeptin cells is a critical mediator of reproductive dysfunction in PCOS. To test this, we utilized Cre-lox technology to conditionally delete androgen receptors from kisspeptin cells (KARKO) in a commonly used mouse model of PCOS induced by using prenatal dihydrotestosterone treatment (prenatal androgen-treated (PNA) mice). To assess estrous cyclicity, vaginal cytology was examined daily for three weeks beginning at 50 days of age. Prenatal vehicle-treated (PNV) wild-type (WT) and PNV KARKO mice (n=6/group) exhibited regular estrous cycles with no significant differences in the percentage of time spent in proestrus, estrus, or metestrus/diestrus, indicating androgen receptors are not required to maintain fertility. As expected, PNA WT mice (n=6) rarely entered proestrus, spent significantly less time in estrus (p<0.05), and spent significantly more time in metestrus and diestrus (p<0.001) compared to the PNV groups (n=6/group, two-way ANOVA), replicating the previously reported acyclic phenotype of PNA mice. No significant differences in the percentage of time spent in cycle stages were detected between PNA WT and PNA KARKO (n=5) mice in this analysis, suggesting androgen signaling at kisspeptin cells is not required to induce acyclicity in PNA mice. Despite this acyclic phenotype, preliminary data using ovarian sections (5μm thickness) stained with hematoxylin and eosin revealed that the percentage of ovarian area composed of corpora lutea was significantly elevated in PNA KARKO mice (n=3) compared to PNA WT mice (n=4) (p<0.05, one-way ANOVA). Further, in this analysis, the area composed of corpora lutea in PNA KARKO mice was not significantly different from PNV WT and KARKO controls (n=6/group), suggesting PNA KARKO mice exhibit a comparable ovulation rate to fertile controls. Together, these data show that androgen signaling at kisspeptin cells partially contributes to the development of anovulation in PNA mice. However, additional mechanisms likely contribute to developing the full PCOS-like reproductive phenotype. Presentation: 6/1/2024

Ovum Pick-Up and Ovaries Characterization of Black Bengal Goat from Slaughterhouse

Black Bengal goat is the only indigenous breed in Bangladesh. These are small-sized goats renowned for their high-quality meat, skin, and adaptability to adverse environmental conditions. To conserve the originality of this species, it is mandatory to develop an effective methodology for their germ cells to be harvested after death. Therefore, the study was conducted to assess the ovarian status and evaluate the most suitable method of ovum pick-up as well as the characterization of ovum from slaughterhouse specimens. A total of 182 ovaries were collected from local slaughterhouses from June 2022 to July 2023 and grossly examined to assess their status. Two methods were used to retrieve oocytes from the ovaries: aspiration and slicing. The grading of the ovum was done based on the germ layer present surrounding the oocytes. From 182 ovaries, the rates of corpus luteum (CL), tumor, and cyst were found to be 14.29%, 6.59%, and 27.47%, respectively. In the aspiration method, the significantly highest percentage of recovery rate (38.89%) and grade A oocytes (51.06%) were found within 1–3 hours of slaughtering. In the slicing method, the significantly highest percentage of recovery rate (19.57%) and grade A (60.61%) oocytes were found within 1–3 hours of slaughtering. The recovery rate was significantly higher (34.1%) in the aspiration method than in the slicing method (17.1%). There was no significant relationship in oocyte quality between the two methods. The recovery rate was not significantly dependent on the presence of CL. In conclusion, the findings from this study may be useful for assessing the quality of oocytes to preserve the germ cells and utilize them for in vitro fertilization for conserving Black Bengal goat.

Reproductive Performance of Ongole Grade, Simmental, and Limousin Cattle Given Superovulation Treatment

Reproductive technologies have the potential to enhance livestock productivity, with superovulation being one such method aimed at increasing the number of ovulated oocytes per estrous cycle through hormonal stimulation. This study aimed to evaluate the reproductive performance of three cattle breeds—Ongole Grade (OG) (n=20), Simmental (n=18), and Limousin (n=20)—subjected to superovulation treatment. The cattle, aged 2.5 to 4 years with a body condition score of 3.5 to 4, were intensively managed. Superovulation was induced using CIDR (Controlled Internal Drug Release) for synchronization, followed by follicle-stimulating hormone (FSH) administration. Reproductive performance data, including age at first estrus, number of corpora lutea (CL), and embryo production, were collected over three estrous cycles. A quasi-experimental design was employed with breed as the primary treatment factor, and data were analyzed using analysis of variance (ANOVA). The results indicated that breed had a significant effect (P0.05) on the age of first estrus, with OG cattle reaching first estrus at 621.3±93.3 days, Simmental at 522.4±75.5 days, and Limousin at 570.4±88.1 days. However, no significant differences (P0.05) were observed among the breeds in terms of the number of CL, collected embryos, or transferable embryos across the superovulation cycles.

The potential role of Ral-interacting protein 76 and vascular endothelial growth factor on angiogenesis in the tumor and ovarian corpus luteum microenvironment.

Tumors and the ovarian corpus luteum have complex mechanisms in the growth microenvironment. Angiogenesis is especially important for demonstrating the molecular mechanism of dynamic cellular function in tumors and corpus luteum. Angiogenesis in tumors and corpus luteum seems to have a similar function, and Ral-interacting protein 76 (RLIP76) and vascular endothelial growth factor (VEGF) are expressed in the tissues of tumors and ovarian corpus luteum. RLIP76 is a potential factor with VEGF in the tumor and corpus luteum angiogenesis. RLIP76 regulates a small GTPase (R-Ras) in cell survival, spreading, and migration. VEGF activates angiogenic functions in tumor and endothelial cells. Hypoxia-inducible factor-1 (HIF-1) is important in tumor growth, tumor angiogenesis, and corpus luteum. VEGF and HIF-1 regulate the angiogenic function of RLIP76, and RLIP76 controls vascular growth in endothelial and tumor cells. RLIP76, R-Ras, VEGF, and HIF-1 may be useful in the research of corpus luteum and cancer therapy and the study of mechanisms of tumor and corpus luteum angiogenesis. This review will help to elucidate the roles of RLIP76 and VEGF in tumor and corpus luteum angiogenesis, tumorigenesis, and the specific regulation of RLIP76 and VEGF. Thus, we reviewed the potential role of RLIP76 and VEGF in the angiogenesis of the tumor and corpus luteum in the tumor and ovarian microenvironment.

Role of ultrasound and color Doppler in diagnosis of ectopic pregnancy and to differentiate from corpus luteum cyst of pregnancy

Background: Ectopic pregnancy is the leading cause of direct maternal deaths. Ectopic pregnancy and corpus luteum cyst are two most common differential diagnoses in a patient with urine pregnancy test (UPT) positive and no sonographic evidence of intrauterine pregnancy. Internal echotexture and color Doppler features of the cyst wall are also considered in an attempt to differentiate the tubal ring of an ectopic pregnancy from that of a corpus luteum. Aims and Objectives: The aim of this study was to diagnose the presence of ectopic pregnancy and to differentiate ectopic pregnancy from corpus luteum cyst using grayscale ultrasound and color Doppler findings. Materials and Methods: The prospective study was conducted in 46 patients who were referred to the Department of Radiodiagnosis in R.L Jalappa Hospital with UPT positive and clinical features suggestive of ectopic pregnancy. Results: Out of 46 patients, 33 were diagnosed with ectopic pregnancy and 13 with corpus luteum cyst. Ectopic pregnancies had thicker walls as compared to corpus luteum cysts. Most of the ectopic pregnancies had hyperechoic walls as compared to ovaries (63.6%) and endometrium (60.6%). Free fluid with echoes was seen in the pelvis in 59.4% ectopic pregnancies whereas most of the corpus luteum cysts (69.2%) had no free fluid. Most of the corpus luteum cysts (61.5%) had clear internal echotexture whereas ectopic pregnancies were mostly lacy or solid. Yolk sac was seen exclusively in ectopic pregnancy (30.3%). RI 0.7 was found to be highly specific for diagnosing ectopic pregnancy. Conclusion: Significant ultrasound parameters that aid in differentiating between the two include the thickness of the mass’s wall, the wall’s echogenicity in relation to the ovary and endometrium, the internal echotexture of the cystic mass, the presence of the yolk sac, and the presence of free fluid with echoes. It was shown that RI 0.7 has a very good specificity in identifying ectopic pregnancy.

Further Insights into The Pathogenic Mechanisms of Haemotropic Mycoplasma ovis.

In this study, we examined the effects of experimental intraperitoneal infection with haemotropic Mycoplasma ovis (0.5 mL of blood containing 80% parasitaemia) on selected serum biomarkers and cellular pathology in mice. After infection, M. ovis cells appeared in the blood films within one week. A dose-dependent peak of parasitemia was observed during the 3rd-week post-infection (pi), with a significant decrease in mean PCV between treatment versus control group at week 3 (t 14 = -3.693, P < 0.02), week 5 (t 14 = -2.096, P = 0.055), and week 7 (t 14 = -4.329, P = 0.001). There was a significantly (t 8 = -2.330, P = 0.048) lower serum oestrogen in treatment (10.38 ± 5.07) than control (17.43 ± 4.48), while serum progesterone was significantly (t 8 = 5.415, P = 0.001) increased in treatment (27.37 ± 2.17) than control (15.92 ± 4.20). Serum haptoglobin was significantly (t 8 = 8.525, P < 0.01) lower in treatment (8.72 ± 1.49) than control (18.16 ± 1.98) while the SAA was significantly (t 8 = 3.362, P = 0.01) higher in treatment (16.79 ± 2.71) than control (11.59 ± 2.15). Prominent lesions observed in the ovary include degeneration, necrosis, vacuolation, and hypertrophy of the lutein cells in corpora lutea. In the lymph nodes, diffused cellular hyperplasia of the lymphoid tissue in the cortex. In the liver, degeneration and necrosis accompanied by leucocytic cellular infiltration and Kupffer cell proliferation within the sinusoids. There were diffused leucocytic infiltrations and proliferative lesions in the glomerulus of the kidneys. The disturbance in progesterone and ovarian pathology highlights the potential role of haemotropic M. ovis in reproductive disorders. The observed changes in biomarkers and cellular reactions following M. ovis infection in the mouse may be further advanced in sheep and goats.

Delaying induction of ovulation and timed AI in a Double-Ovsynch protocol increased expression of estrus and altered first service reproductive outcomes of lactating dairy cows

The objective of this randomized controlled experiment was to evaluate the effect of delaying induction of ovulation and timed artificial insemination (TAI) on expression of estrus before AI and first service reproductive outcomes. A secondary objective was to evaluate the effects of delaying induction of ovulation in a Double-Ovsynch protocol on ovarian function. Lactating Holstein cows (n = 4,672) from 2 commercial dairy farms fitted with sensors for automated detection of estrus were synchronized with a Double-Ovsynch protocol up to the first PGF2α (PGF-L) of the Breeding-Ovsynch portion of the protocol (Pre-Ovsynch: GnRH, 7 d later PGF2α, 3 d later GnRH, 7 d later Breeding-Ovsynch: GnRH, 7 d later PGF2α, 1 d later PGF2α). At PGF-L, cows blocked by parity (primiparous vs. multiparous) and semen used for first service (sex-sorted dairy vs. conventional beef) were randomly assigned to the G56 (n = 2,338) or G80 (n = 2,334) treatments. Cows in G56 had 56 h whereas cows in G80 had 80 h from PGF-L to induction of ovulation with the last GnRH (GnRH2) before AI. For both treatments, TAI occurred ∼16 h after GnRH2. All cows with automated estrus alerts between PGF-L and TAI were inseminated at detected estrus (AIE) without GnRH. Ovarian function and responses to synchronization were monitored based on circulating concentrations of progesterone and examination of the ovaries by ultrasonography. Data for binary outcomes were analyzed by logistic and continuous outcomes with lineal regression. More cows in G80 received AIE and had estrus before AI. Overall, pregnancies per AI (P/AI) did not differ for the G80 and G56 treatments. Cows in G80 that received TAI and had no estrus had fewer P/AI than cows with estrus that received AIE or TAI in G80, and fewer P/AI than cows AIE and cows that received TAI and had or did not have estrus in the G56 treatment. No differences were observed between treatments or for cows with and without estrus for pregnancy loss. Unlike some minor differences between treatments for concentrations of progesterone at GnRH2, the most notable differences in ovarian function were for cows in both treatments with or without estrus that received TAI. Cows with estrus, were more likely to have follicles > 16 mm, had larger follicles before ovulation, and had a greater ovulation risk after AI. Likewise, within the G80 treatment only, cows with estrus that received AIE or TAI had larger follicles, were more likely to have complete luteal regression, had greater ovulation risk, were more likely to have a functional corpus luteum, and had more circulating progesterone after AI. We concluded that delaying induction of ovulation and TAI was effective for allowing more cows to express estrus before AI which had different ovarian function outcomes and greater P/AI than cows that did not express estrus. However, the greater P/AI of cows that expressed estrus was insufficient to compensate for the reduced P/AI of cows that did not express estrus, and thus increase overall P/AI compared with the treatment without delayed induction of ovulation. Detection of estrus before AI in cows undergoing synchronization of ovulation could help identify cows with different likelihoods of pregnancy after insemination.

Structural basis of selective beta-carotene binding by a soluble protein

β-carotene (BCR) is the most abundant carotenoid, a colorant, antioxidant, and provitamin A. The extreme hydrophobicity of this hydrocarbon requires special mechanisms for distribution in aqueous media, including water-soluble carotenoproteins. However, all known carotenoproteins prefer oxygenated carotenoids and bind BCR inefficiently. Here, we present the crystal structure of the BCR-binding protein (BBP) from gregarious male locusts, which is responsible for their vivid yellow body coloration, in complex with its natural ligand, BCR. BBP forms an antiparallel tubular homodimer with α/β-wrap folded monomers, each forming a hydrophobic 47Å long, coaxial tunnel that opens outward and is occupied by one s-cisC6-C7, all-trans BCR molecule. In the BCR absence, BBP accepts a range of xanthophylls, with reduced efficiency depending on the position and number of oxygen atoms, but rejects lycopene. The structure captures a pigment complex with a Takeout 1 protein and inspires potential applications of BBP as a BCR solubilizer.

The Effects of L-Tartaric Acid on Ovarian Histostereological and Serum Hormonal Analysis in an Animal Model of Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) is the most common endocrine-related reproductive disorder in women of reproductive age, accompanied by both the impairment of female fecundity and a risk of metabolic disorders. PCOS is emphasized as a worldwide concern due to its unknown etiology and lack of specific medications. The current study aimed to evaluate the effects of L-tartaric acid, an abundantly occurring compound in fruits, on the histostereological and hormonal changes caused by PCOS. Forty adult Sprague Dawley rats were randomly divided into four groups including controls (no intervention), Tartaric acid (40mg/Kg/day from day 21 onwards for 39 days), PCOS (21 days letrozole and then normal saline orally for 39 days), and PCOS + Tartaric acid. After treatments, the ovarian histostereological analysis as well as the level of reproductive hormones including luteinizing hormone (LH), follicle-stimulating hormone (FSH), estradiol, progesterone, and testosterone was measured. PCOS caused a significant decrease in the number of unilaminar, multilaminar, antral, and graafian follicles and increased follicular atresia (p-value < 0.001). Moreover, the weight and volume of ovarian tissue and related structures including cortex, medulla, and cysts increased significantly (p-value < 0.0001). However, corpus luteum volume was significantly decreased (p-value < 0.001). Although significant differences were found in some parameters with the control group (p-value < 0.05), the administration of tartaric acid restored the pathological effects of PCOS on the ovarian histostructure. Furthermore, tartaric acid improved the serum levels of LH, estradiol, progesterone, and testosterone (p-value < 0.05). The obtained findings may suggest tartaric acid as a novel strategy for PCOS management, although further studies are necessary.

Neuregulin 1 Signaling Attenuates Tumor Necrosis Factor α-Induced Female Rat Luteal Cell Death.

The corpus luteum (CL) is a transient ovarian endocrine structure that maintains pregnancy in primates during the first trimester and in rodents during the entire pregnancy by producing steroid hormone progesterone (P4). CL lifespan, growth, and differentiation are tightly regulated by survival and cell death signals through luteotrophic and luteolytic factors, including the epidermal growth factor (EGF)-like factor family. Neuregulin 1 (NRG1), a member of the EGF family, mediates its effect through ErbB2/3 receptors. However, the functional role of NRG1 in luteal cells (LCs) is unknown. Thus, this study investigated the role of NRG1 and its molecular mechanism of action in rat LC. Our experimental results suggest a strong positive correlation between steroidogenic acute regulatory protein (StAR) and NRG1 expression in mid-CL and serum P4 and estrogen (E2) production. In contrast, there was a decrease in StAR and NRG1 expression and P4 and E2 production with an increase in tumor necrosis factor α (TNFα) expression in regressing CL. Further in vitro studies in LCs showed that the knockdown of endogenous Nrg1 promoted the expression of proinflammatory and proapoptotic factors and decreased prosurvival factor expression. Subsequently, treatment with exogenous TNFα under these experimental conditions profoundly elevated proinflammatory and proapoptotic factors. Further analysis demonstrated that the phosphorylation status of ErbB2/3, PI3K, Ak strain transforming or protein kinase B (Akt), and ErK1/2 was significantly inhibited under these experimental conditions, whereas the treatment of TNFα further inhibited the phosphorylation of ErbB2/3, PI3K, Akt, and ErK1/2. Collectively, these studies provide new insights into the NRG1-mediated immunomodulatory and prosurvival role in LCs, which may maintain the function of CL.

Lactylation drives hCG-triggered luteinization in hypoxic granulosa cells

Hypoxia that occurs during the luteinization process of granulosa cells (GC) contributes to the formation of lactate in follicles. Lysine lactylation (Kla), a post-translational modification directly regulated by lactate levels, is a metabolic sensor that converts metabolic information into gene expression patterns. In this study, we employed human chorionic gonadotropin (hCG) to induce GCs luteinization and discovered that hypoxia enhances hCG-mediated GCs luteinization by stimulating lactate production/lactylation. The elevated levels of luteinization markers (including progesterone synthesis, expression of CYP11A1 and STAR) were accompanied by increased lactate production as well as enhanced lactylation in mouse ovarian GCs after the injection of hCG in vivo. By treating GCs with hypoxia in vitro, we found that hypoxia accelerated hCG-induced GCs luteinization, which was inhibited after blocking lactate production/lactylation. Further investigations revealed that H3K18la might contribute to hCG-induced luteinization in hypoxic GCs by upregulating CYP11A1 and STAR transcription. Additionally, we identified that CREB K136la is also required for hCG-induced GCs luteinization under hypoxia. Finally, the in vitro findings were verified in vivo, which showed impaired GCs luteinization and corpus luteum formation after blocking the lactate/lactylation by intraperitoneal injection of oxamate/C646 in mice. Taken together, this study uncovered a novel role of protein lactylation in the regulation of GCs luteinization.